Quantitative Queries
How many moles are in 20g of NaCl (MW: 58g/mol)?
0.345 moles
Incubating the SDS Page protein samples at high temperatures is designed to help further denature the proteins by breaking these types of bonds.
What are Hydrogen bonds?
There is an inverse relationship between enzyme reaction rates and this value
What is the Michaelis constant (Km)?
“Molarity” and “mg/mL” are measures of _________ of solutions, while “mg” and “moles” are measure of ________ of solutes.
What is concentration; amounts?
This graphical representation linearizes the Michaelis-Menten equation, allowing for the determination of kinetic parameters Km and Vmax through the inverse of substrate concentration and reaction velocity.
What is a Lineweaver-Burk plot?
How many milligrams are in 18 moles of NaOH? (MW: 40g/mol)
720,000 mg
This anionic detergent is commonly used in gel electrophoresis to denature proteins and impart a negative charge, ensuring that they separate based solely on their size rather than shape or charge.
What is SDS (Sodium Dodecyl Sulfate)
When all the optimum environmental conditions needed for enzyme action are met, the velocity of an enzyme catalyzed reaction is under the control of these two factors
what is enzyme and substrate concentrations
After running a gel, this visualization technique, which involves staining the gel a bright blue color to highlight proteins, is crucial for determining the number of bands present and assessing protein purity.
What is Coomassie staining?
This term refers to the maximum rate of reaction that an enzyme can achieve when fully saturated with substrate.
What is Vmax?
Given a 2mM stock, you are asked to make 10 mL of a a 5uM solution. How many mL of the stock do you need?
0.025 mL
In SDS-PAGE, as proteins migrate from the cathode to the anode, their migration rate decreases as this quantity increases, leading to size-based separation.
What is the molecular weight of the protein?
These non-protein molecules are required for the catalytic activity of some enzymes and can be metal ions or organic molecules.
What are cofactors or coenzymes?
This type of light, consisting of a single wavelength, is essential in measuring the concentrations of substances using Beer's Law, reducing interference, and ensuring precise absorbance measurements during the monitoring of enzymatic reactions.
What is monochromatic light?
The value of Km provides insight into this aspect of the enzyme’s interaction with its substrate, and a low Km suggests this type of affinity between the enzyme and substrate.
What is a high affinity?
Given 90% solution of ethanol, what volume would you need to make 50mL of a 65% ethanol solution?
36.1 mL
In gel electrophoresis, the Rf value is influenced by factors such as the gel concentration and the size of the molecules; as the size increases, this typically happens to the Rf value.
What causes a decreasing Rf value?
Michaelis and Menten found that when the enzyme is held constant while the amount of S is varied. This graph relationship between velocity and substrate concentration is observed.
What is hyperbolic?
This concept, crucial for Michaelis-Menten kinetics, assumes that at enzyme-substrate saturation, the formation and breakdown of the enzyme-substrate complex reach a balance, leading to a constant reaction rate despite increasing substrate concentration
What is steady-state equilibrium?
This law describing a mathematical relationship is often represented in enzyme kinetics using a standard curve, where the slope indicates the molar absorptivity, allowing for the quantification of substrate conversion based on absorbance readings.
What is Beer's Law (or the Beer-Lambert Law)?
If you have a stock solution of 10 M, how many 1:10 serial dilutions would you need to achieve a final concentration of 100 uM?
5
This property of amino acids allows Coomassie Brilliant Blue (Bradford Reagent) to bind effectively, enhancing protein concentration quantitation.
What is the positive charge?
In the presence of this type of inhibitor, Vmax remains unchanged, but the apparent Km increases due to competition between the substrate and inhibitor for the enzyme's active site.
What is a competitive inhibitor?
At steady state equilibrium of enzyme catalyzed reaction, the rate-determining step of the reaction rate per Michaelis Menten kinetics is
What is k3?
In studying enzyme kinetics for an enzyme catalyzed reaction, the slope of line relating the amount of product produced over a period of time is
What is initial velocity?